抗原

Get high purity DCs by Cultured plastic-adherent monocytes isolated from healthy human peripheral blood with GM-CSF and IL-4 for 7 days. To observe the morphology of DCs by inverted phase contrast microscope ,electron microscope and laser confocal microscope. Analyse phenotype of DCs with flow cytometry. Investigate the endocytosis ability of DCs as a group by Horseradish peroxidase endocytosis assay. To appraise allogeneic mixed lymphocytes reaction of DCs by MTT reduction assay. Analyse the levels of IL-12 and TNF in liquids of cultured medium by ELISA and MTT reduction assay respectively. Soluble antigens of HCCs was obtained by 3 freeze-and-thaw cycles. Biological characteristics of HC soluble antigens pulsed DCs were monitored by flow cytometry. According to MTT reduction assay estimated the cell proliferation of self lymphocytes activated by HC antigens pulsed DCs. Get high purity BCG HSP 70 protein by SDS-PAGE electrophoresis and determined its biological activity with ELISA. Analyse phenotype of antigen pulsed DCs primed by BCG HSP70 with flow cytometry. By MTT reduction assay estimated the cell proliferation of self lymphocytes and the MLR of DC based vaccine. Analyse expression of HLA-DR molecule on surface of HCC lines. The IFN-γ mRNA in lymphocytes after actived by DC vaccine and the Fas-L expression on DC and DC vaccine primed lymphocytes were detected by in situ hybridization and flow cytometry respectively. Specific cytotoxity lysis of T lymphocytes and nonspecific inhibition of liquids in culture medium against HCC lines were also tested. Detect expression of hAFP on four HCC lines with Cell-ELISA. Induce apoptosis of HCCs with actinomycin-D. Interaction of DCs and apoptotic cells was observed under transmission electron microscope. Growth inhibition test of DC against HCC lines was also performed. Establish the nude mouse model bearing human HC xenografts and indentify the characteristic of tumour by histochemistry and immunohistochemistry techniques. Prevent and treat transplanted human HC on nude mouse with Freezing and anabiotic HC specific lymphocytes.

用GM-CSF和IL-4从健康人外周血诱导DC；分别用倒置相差显微镜、电子显微镜及激光共聚焦显微镜观察DC形态；流式细胞术检测DC表型；HRP吞噬实验测定DC的群体内吞能力；MTT法检测同种异体混合淋巴细胞反应；ELISA法和MTT法分别测定DC培养上清液中IL-12和TNF水平；冻融法制备肝癌细胞可溶性抗原；流式细胞术检测负载肝癌可溶性抗原后DC的生物学特性；MTT法检测DC负载肝癌抗原后对自身淋巴细胞增殖的影响；SDS-PAGE制备电泳纯化BCG HSP70并鉴定纯度，ELISA测定活性；流式细胞术检测负载抗原DC经BCGHSP 70活化后的表型；MTT法检测肝癌DC疫苗对自身淋巴细胞增殖的影响和混合淋巴细胞反应；流式细胞术检测肝癌细胞表面HLA-DR表达；MTT法检测肝癌DC疫苗对自身淋巴细胞的活化；原位杂交法检测肝癌DC疫苗活化后的淋巴细胞IFN-γmRNA表达；流式细胞术检测DC和肝癌DC疫苗活化后淋巴细胞表面Fas-L；MTT法分别检测肝癌DC疫苗活化的淋巴细胞和其培养上清对肝癌细胞的特异性杀伤和非特异性抑制作用；Cell-ELISA检测人肝癌细胞hAFP表达；MTT法检测负载AFP表位肽和凋亡肝癌细胞DC对自身淋巴细胞增殖的影响；ELISA法和MTT法分别测定活化后淋巴细胞培养上清中TNF和IL-12水平；肝癌细胞凋亡的诱导和检测；DC吞噬凋亡肝癌细胞后的电子显微镜观察；DC对肝癌细胞的生长抑制试验；人肝癌裸鼠皮下移植瘤动物模型的建立及其组织学和免疫组织化学鉴定；DC及肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤；冻存和复苏后的肝癌特异性淋巴细胞预防和治疗人肝癌裸鼠皮下移植瘤。

ResultsThe diagnostic sensitivity of ELISA using excretory-secretory antigen was 92.5%, which was higher than that of ELISA using crude Clonorchis sinensis antigen(88.2%).

结果比较华支睾吸虫分泌-排泄抗原的诊断的敏感性高于粗抗原，分泌-排泄抗原的特异性反应明显高于粗抗原。

Theoretically, its calculation complexity is studied, while it is tested empirically by learning multi-samples generated randomly. The comparative analysis and applications show it has some merits of rapid searching and good clustering effect. B3. An immune network optimization algorithm with information feedback, which can solve special complex function optimization problems with high dimensions which objective functions are formed by summing multi-subobjective ones, is constructed based on the clone selection principle and the immune network regulation of the idiotype immune network.

为了对免疫系统中抗体如何学习抗原特征的整体行为有一定的认识，从单个抗体识别抗原及多个抗体并行识别抗原的特征出发，利用免疫应答的相关原理及免疫细胞的免疫功能设计了适合于批量样本聚类的抗原特征学习算法，理论上分析了其计算复杂度，获得了此算法中的关键参数与其搜索性能的关系；应用上，该算法通过对实际的多样本学习，获得了搜索速度快及聚类效果好的优点，比较分析表明这种方法是可行的和有效的。

Following attachment to the host immunized with purified antigens, midgut antigens, salivary gland antigens and reinfestation by I. sinensis, the midgut of tick revealed rather strikingly pathological changes, especially of the tick feeding on host immunized with 105KD purfied antigens the basal lamina became thinner, looser, and broken.

中华硬蜱叮咬纯化抗原、中肠抗原、唾液腺抗原免疫接种组和再次感染组宿主后，中肠可发生一系列明显的病理变化，尤以叮咬105KD纯化抗原组宿主后中肠病理变化最为严重。

The fused dominant peptide antigen 38 kD-ESAT6-CFP10 might serve as a serodiagnostic candidate antigen, and the double antigen sandwich ELISA using the fused antigen might be used to detect the tuberculosis and improve the specificity and sensitivity of diagnostic reagent.

该种抗原优势肽段融合抗原有望成为结核病血清学诊断的重要候选抗原，并且双抗原夹心ELISA方法有助于提高检测的特异性和敏感性。

In this research, B2 gene of HCV was highly expressed as a fusion protein (Trx-B2) by cloning into pThioHisA, and induced specific anti-HCV antibody in higher titer than unmodified B2 protein after immunize mice or rabbit.The results showed antigen Trx-B2 has obvious immunogenicity and induced specific antibody,which might be able to serve as diagnosis HCV. Consequently, this PcAb of rabbit was used to coated Immulon plate, and a method of both antibodys sandwiching antigens was developed to detect 32 HCV positive serum and 32 nomal serum.Results show that ratio of positive are 87.5%and 43.75% respectively.

但缺点是该抗原所诱导的抗体滴度较低，可能是由于其分子量较小，所以本课题将人工合成的HCV复合多表位抗原基因B_2克隆到表达载体pThioHisA中，利用硫氧还蛋白融合方式对该抗原进行修饰，并在大肠杆菌中表达融合蛋白(Trx-B_2)，纯化该蛋白后免疫小鼠及家兔，结果表明用硫氧还蛋白修饰后的融合蛋白免疫小鼠和家兔后，得到的抗体滴度均高于未经修饰过的复合多表位抗原合成肽所诱导的抗体滴度，并从免疫后的兔血清中提取IgG包被酶联板，利用双抗夹心法分别检测了32份HCV阳性血清和32份正常人的血清，结果阳性率分别为87.5%和43.75%，符合率为71.88%。

Methods Indirect immnofluorescence assay was applied to identify specificity of the two groups of monoclonal antibodies prepared with crude antigen and recombinant antigen of Aspergillus fumigatus,respectively.Two different double monoclonal antibody sandwich ELISA assays established with the two groups of antibodies were performed to detect antigents in the cell culture supermatants of 19 common species of Aspergillus,Penicillium marneffei,and 5 species of Candidas.

采用天然烟曲霉抗原免疫，获得广谱针对曲霉抗原的单克隆抗体；采用重组烟曲霉抗原获得特异性针对烟曲霉抗原的单克隆抗体，用间接免疫荧光鉴定，并分别建立2种双抗体夹心ELISA法，对19种常见的环境和临床分离曲霉株、马尔尼菲氏青霉菌及念珠菌培养液进行检测。

Two types of haptens, the earboxylated propyl amino derivative of the acid moiety (HCA1) that is the hydrolyzed product of cyhalothrin, and the carboxylated ethyl carbonyl derivative of the alcohol moiety (HCB2) were synthesized, and conjugated with the carrier proteins bovine serum albumin and ovalbumin respectively by the active ester method and the mixed anhydride method. The polyclonal antibodies were obtained after immuning to the New Zealand rabbits.

通过对氯氟氰菊酯分子结构中的酸部分接4-氨基丁酸(HCA1)和醇部分接丁二酸酐(HCB2)，合成两种不同的半抗原，并通过碳二亚胺法将HCA1和HCB2分别与牛血清蛋白偶联制备了人工抗原HCA1-BSA和HCB2-BSA，通过活泼酯法将HCA1、HCAS和HCB2分别与卵清蛋白偶联制备包被抗原，两种人工抗原通过免疫新西兰大白兔来制备相应的多克隆抗体。

Our preliminary results come to the conclusions that:(1)there were more common antigens or epitopes of common antigens in oviducts of different mammalian animals;(2)the antibodies against porcine and bovine ovidu...

提示人和猪以及人和牛输卵管抗原中存在较多类似的抗原决定簇。鉴于透明带抗原有卵巢源和输卵管源，因此本研究结果为人和猪透明带抗原中存在着明显交叉性提拱了部分依据。

Pneumophila serogroup 5 antibody to develop two immuo-colloidal gold tests respectively. Main works listed below as:1、Preparation and identification of polysaccharide antigen of L. pneumophila Bacteria of LP1 ~ 7,9 and 10 were cultured on yeast agar buffer activated carbon for three to five days at 37℃, 5%CO2. Protein-free polysaccharide antigens were obtained after harvest in cells, extraction, deproteinization, dialysis and other steps. Their immunogenicities were verified by ultraviolet spectrophotometer full-wavelength scanning and Western blotting.2、Preparation and identification of rabbit anti-LP1 antibodies and rabbit anti-LP5 antibody Rabbit anti-LP1 and anti-LP5 antibodies were purified after rabbits were immuned with antigens isolated as described above. The purities of both antibodies were above 80% and the titer of blood serum 1:32 tested by double antibody sandwich assay.3、Development of colloidal gold immunochromatographic assay kit The size of colloidal gold particles in the kit was 25nm. The optimal concentrations for antibodies were 30μg/ml and the sensitized concentrations of NC membrane were 5 mg/ml.

主要研究工作从以下几个方面进行：1、LP1~7、9和10型多糖抗原的制备与鉴定将LP1~7、9和10型菌株分别接种在缓冲活性炭酵母琼脂培养基上，37℃、5%CO2的条件下培养，3~5天后洗下菌苔，经抽提、除蛋白、透析等步骤后得到基本无蛋白的LP多糖抗原，经紫外分光光度仪全波长扫描及Western blotting验证其抗原良好。2、兔抗LP1抗体和兔抗LP5抗体的制备与鉴定分别用LP1、LP5型多糖抗原免疫家兔，采用琼脂糖双向扩散试验检测，两种抗体血清效价均为1：32；饱和硫酸铵法提取抗体，SDS-PAGE检定其抗体纯度均达到80%以上。3、胶体金免疫层析检测试剂的初步研制采用柠檬酸三钠还原法制备约25nm大小的胶体金颗粒；分别制备兔抗LP1抗体、兔抗LP5抗体的金标探针，两种抗体的最适标记量均为30μg/ml；选择适当孔径的微孔滤膜为载体包被两种抗体，NC膜包被浓度均为5mg/ml。

As she looked at Warrington's manly face, and dark, melancholy eyes, she had settled in her mind that he must have been the victim of an unhappy attachment.

每逢看到沃林顿那刚毅的脸，那乌黑、忧郁的眼睛，她便会相信，他一定作过不幸的爱情的受害者。

Maybe they'll disappear into a pothole.

也许他们将在壶穴里消失